POWRE: Role of Calcium-Binding Proteins in Synaptic Transmission Between Retinal Amacrine Cells

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This POWRE Research Enhancement award will give Dr. Gleason the opportunity to explore the feasibility of using antisense techniques to investigate her research system of retinal neurons. Control of calcium concentration in cells is critical for regulation of calcium-dependent events such as synaptic transmission between neurons. Calcium homeostasis is especially important in the retina, where synaptic signaling is relatively slow and therefore calcium elevations can be large. This research project addresses the role that buffering calcium-binding proteins play in neuronal calcium homeostasis and synaptic transmission. Dr. Gleason will use antisense oligonucleotides to block the expression of the predominant buffering calcium-binding protein, calretinin, then examine the effects of removing this protein on synaptic transmission. Synaptic transmission will be studied in a preparation of cultured neurons derived from the chick retina, where an identified type of retinal neuron, the amacrine cell, forms synapses with other amacrine cells. Isolated pairs of synaptically connected amacrine cells can be studied with high resolution using simultaneous perforated-patch voltage-clamp recording of both neurons. By combining the two powerful techniques of molecular biology and electrophysiology, this research will enhance our understanding of calcium-homeostasis mechanisms in the central nervous system. The antisense methods developed in this pilot project will be used in future studies to selectively remove other neuronal proteins that may be important in retinal synaptic transmission.